The appearance and spread of antibiotic-resistant pathogens known as antimicrobial resistance(AMR)is one of the major worldwide health crises that humanity have to deal with over the next decades.One of the main metho...The appearance and spread of antibiotic-resistant pathogens known as antimicrobial resistance(AMR)is one of the major worldwide health crises that humanity have to deal with over the next decades.One of the main methods for addressing AMR is the effective screening for antimicrobial insensitivity in clinical and environmental monitoring.Current clinical laboratory procedures use traditional culturebased antibiotic susceptibility testing(AST)methods,which can take up to 24 h to identify which drug is suitable for the infection inhibition.Therefore,it is vital to develop novel strategies that offer quick,simple,affordable,reliable,sensitive and accurate AMR monitoring.Sensors for AMR markers detection could possess the essential qualities for quickly identifying resistant microorganisms and could give vital data for the selection of antibacterial drugs administration.This review offers a summary of the innovative application of these AMR markers detection strategies focusing on healthcare and environmental surveillance for the AMR genotypic or phenotypic assessment.展开更多
BACKGROUND: There are 8 well-documented genotypes of hepatitis B virus (HBV) at this time point. Genotyping can be accomplished based on a partial sequence of hepatitis B virus (HBV) genome such as the pre-S or S gene...BACKGROUND: There are 8 well-documented genotypes of hepatitis B virus (HBV) at this time point. Genotyping can be accomplished based on a partial sequence of hepatitis B virus (HBV) genome such as the pre-S or S gene. Several methods have been developed and used for HBV genotyping including direct sequencing, restriction fragment length polymorphism, line probe assay and enzyme-linked immunoassay. Recently, a novel, rapid and cost-effective genotyping method based on PCR amplification assay using type-specific primers that can identify all six major genotypes has been developed. This study was undertaken to characterise HBV genotypes and investigate the association between the prevalence of different genotypes and the severity of HBV-induced liver diseases. METHODS: Serum samples from carriers of HBV and patients with HBV-related liver diseases from Zhejiang Province were screened for viral serological markers using commercially available radioimmunoassay (RIA) and enzyme linked immunosorbent assay (ELISA) kits. Serum HBV DNA load was determined by real-time detection PCR. A type-specific primer based the nested-PCR method was employed in the HBV genotyping. The genotype results obtained were confirmed by direct sequencing of nested PCR amplicons of the pre-S region. Ten samples of each genotype (B and C) were sequenced. RESULTS: The survey on a cohort of 125 HBV carriers in and around Hangzhou City, Zhejiang Province showed the existence of HBV genotypes A (0.8%), B (48%), C (40.8%), D (0.8%), mixed B and C (9.6%) and an absence of E and F genotypes. Distribution of HBV genotypes in patients with liver diseases revealed a statistically insignificant higher prevalence of genotype B in mild chronic hepatitis (CH). Among the three genotypes B, C and mixed B/C infections 11 (73.3%), 3 (20%) and 1 (6.7%), (P< 0.05), respectively in subjects with moderate CH, genotype B was significantly predominant. The infection patterns for genotypes B, C and B/C mixed in (i) liver cirrhosis (LC) 4 (23.5%), 10 (58.8%) and3 (17.7%)展开更多
苹果蠹蛾Cydia pomonella(L.)是我国重大农业入侵害虫,对我国果业健康发展造成严重威胁。在利用基因编辑等技术对相关基因进行功能研究时,通常采用单对交配策略对纯合突变体进行筛选。因此,在配对前明确个体基因型,避免对虫体造成损伤...苹果蠹蛾Cydia pomonella(L.)是我国重大农业入侵害虫,对我国果业健康发展造成严重威胁。在利用基因编辑等技术对相关基因进行功能研究时,通常采用单对交配策略对纯合突变体进行筛选。因此,在配对前明确个体基因型,避免对虫体造成损伤尤为重要。本研究分别收集末龄幼虫蜕(10个蜕)和蛹壳(1个、5个、10个、15个蛹壳),通过基因组DNA提取、目的基因PCR扩增、琼脂糖凝胶电泳检测和PCR产物测序,以评估该方法作为苹果蠹蛾基因检测的可行性。结果显示,以苹果蠹蛾末龄幼虫10个蜕及5个以上蛹壳提取的基因组DNA作为模板,扩增精巢特异性丝氨酸/苏氨酸蛋白激酶(Testis specific serine/threonine protein kinase,TSSK1)基因,PCR产物经琼脂糖凝胶电泳检测得到单一、明亮的条带,经测序证实该产物是目的基因序列。基于高效及节约成本的原则,拟推荐利用10个末龄幼虫蜕或10个蛹壳提取基因组DNA、通过PCR扩增和测序进行苹果蠹蛾的无损伤基因型检测。本研究建立了一种利用幼虫蜕和蛹壳进行苹果蠹蛾无损伤、高效的基因检测方法,为提高苹果蠹蛾基因功能研究的效率奠定了基础。展开更多
基金supported by the National Natural Science Foundation of China(No.82202221)the Natural Science Foundation of Anhui Province(No.2208085QB39)College Students’Innovative Entrepreneurial Training Plan Program(No.202110359071).
文摘The appearance and spread of antibiotic-resistant pathogens known as antimicrobial resistance(AMR)is one of the major worldwide health crises that humanity have to deal with over the next decades.One of the main methods for addressing AMR is the effective screening for antimicrobial insensitivity in clinical and environmental monitoring.Current clinical laboratory procedures use traditional culturebased antibiotic susceptibility testing(AST)methods,which can take up to 24 h to identify which drug is suitable for the infection inhibition.Therefore,it is vital to develop novel strategies that offer quick,simple,affordable,reliable,sensitive and accurate AMR monitoring.Sensors for AMR markers detection could possess the essential qualities for quickly identifying resistant microorganisms and could give vital data for the selection of antibacterial drugs administration.This review offers a summary of the innovative application of these AMR markers detection strategies focusing on healthcare and environmental surveillance for the AMR genotypic or phenotypic assessment.
文摘BACKGROUND: There are 8 well-documented genotypes of hepatitis B virus (HBV) at this time point. Genotyping can be accomplished based on a partial sequence of hepatitis B virus (HBV) genome such as the pre-S or S gene. Several methods have been developed and used for HBV genotyping including direct sequencing, restriction fragment length polymorphism, line probe assay and enzyme-linked immunoassay. Recently, a novel, rapid and cost-effective genotyping method based on PCR amplification assay using type-specific primers that can identify all six major genotypes has been developed. This study was undertaken to characterise HBV genotypes and investigate the association between the prevalence of different genotypes and the severity of HBV-induced liver diseases. METHODS: Serum samples from carriers of HBV and patients with HBV-related liver diseases from Zhejiang Province were screened for viral serological markers using commercially available radioimmunoassay (RIA) and enzyme linked immunosorbent assay (ELISA) kits. Serum HBV DNA load was determined by real-time detection PCR. A type-specific primer based the nested-PCR method was employed in the HBV genotyping. The genotype results obtained were confirmed by direct sequencing of nested PCR amplicons of the pre-S region. Ten samples of each genotype (B and C) were sequenced. RESULTS: The survey on a cohort of 125 HBV carriers in and around Hangzhou City, Zhejiang Province showed the existence of HBV genotypes A (0.8%), B (48%), C (40.8%), D (0.8%), mixed B and C (9.6%) and an absence of E and F genotypes. Distribution of HBV genotypes in patients with liver diseases revealed a statistically insignificant higher prevalence of genotype B in mild chronic hepatitis (CH). Among the three genotypes B, C and mixed B/C infections 11 (73.3%), 3 (20%) and 1 (6.7%), (P< 0.05), respectively in subjects with moderate CH, genotype B was significantly predominant. The infection patterns for genotypes B, C and B/C mixed in (i) liver cirrhosis (LC) 4 (23.5%), 10 (58.8%) and3 (17.7%)
文摘苹果蠹蛾Cydia pomonella(L.)是我国重大农业入侵害虫,对我国果业健康发展造成严重威胁。在利用基因编辑等技术对相关基因进行功能研究时,通常采用单对交配策略对纯合突变体进行筛选。因此,在配对前明确个体基因型,避免对虫体造成损伤尤为重要。本研究分别收集末龄幼虫蜕(10个蜕)和蛹壳(1个、5个、10个、15个蛹壳),通过基因组DNA提取、目的基因PCR扩增、琼脂糖凝胶电泳检测和PCR产物测序,以评估该方法作为苹果蠹蛾基因检测的可行性。结果显示,以苹果蠹蛾末龄幼虫10个蜕及5个以上蛹壳提取的基因组DNA作为模板,扩增精巢特异性丝氨酸/苏氨酸蛋白激酶(Testis specific serine/threonine protein kinase,TSSK1)基因,PCR产物经琼脂糖凝胶电泳检测得到单一、明亮的条带,经测序证实该产物是目的基因序列。基于高效及节约成本的原则,拟推荐利用10个末龄幼虫蜕或10个蛹壳提取基因组DNA、通过PCR扩增和测序进行苹果蠹蛾的无损伤基因型检测。本研究建立了一种利用幼虫蜕和蛹壳进行苹果蠹蛾无损伤、高效的基因检测方法,为提高苹果蠹蛾基因功能研究的效率奠定了基础。
